8d4d

gamma-Arf1 mediated dimeric assembly of AP-1, Arf1, Nef complex within lattice on MHC-I lipopeptide incorporated narrow membrane tubes

Method: ELECTRON MICROSCOPY Dmax: 219.8 Å Quality: GOOD

1. Protein Identity and Related Structures Protein Identity & Related Structures

ADP-ribosylation factor 1

Homo sapiens

UniProt P84077

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 18 PDB declaration: octadecameric(18) Consistent with protein copy count Chain C; UniProt 2–181 Chain D; UniProt 2–181 Chain F; UniProt 2–181 Chain H; UniProt 2–181 Not recorded Protein Nef × 4 (Q90VU7) HLA class I histocompatibility antigen, A alpha chain × 2 (P04439) AP-1 complex subunit beta-1 × 2 (Q10567) AP-1 complex subunit gamma-1 × 2 (P22892) AP-1 complex subunit mu-1 × 2 (P35585) AP-1 complex subunit sigma-3 × 2 (Q96PC3) GTP GUANOSINE-5'-TRIPHOSPHATE × 4 MG MAGNESIUM ION × 4 ELECTRON MICROSCOPY cryo-EM buffer:pH 7.2;HEPES/KOAc concentrated stocks are diluted to their final concentrations then pH'd to 7.2 with KOH prior to use in experiments. cryo-EM vitrification conditions:Cryogen ETHANE;60 second wait, 3-5 second blot, 597 filter paper, 0.5 second drain. Sample was supplemented with 10nm BSA-gold fiducials. 3.5ul of the mixture was double-side blotted. Resolution 9.60 Å

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

39 other PDB entries and 46 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name ARF1_HUMAN
Isoform
PDB entities 1
Chains and sequence ranges Author chain C; PDBConstruct 1–180; UniProt 2–181 Author chain D; PDBConstruct 1–180; UniProt 2–181 Author chain F; PDBConstruct 1–180; UniProt 2–181 Author chain H; PDBConstruct 1–180; UniProt 2–181

Protein Nef

Human immunodeficiency virus 1

UniProt Q90VU7

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 18 PDB declaration: octadecameric(18) Consistent with protein copy count Chain I; UniProt 2–206 Chain K; UniProt 2–206 Chain L; UniProt 2–206 Chain N; UniProt 2–206 Not recorded ADP-ribosylation factor 1 × 4 (P84077) HLA class I histocompatibility antigen, A alpha chain × 2 (P04439) AP-1 complex subunit beta-1 × 2 (Q10567) AP-1 complex subunit gamma-1 × 2 (P22892) AP-1 complex subunit mu-1 × 2 (P35585) AP-1 complex subunit sigma-3 × 2 (Q96PC3) GTP GUANOSINE-5'-TRIPHOSPHATE × 4 MG MAGNESIUM ION × 4 ELECTRON MICROSCOPY cryo-EM buffer:pH 7.2;HEPES/KOAc concentrated stocks are diluted to their final concentrations then pH'd to 7.2 with KOH prior to use in experiments. cryo-EM vitrification conditions:Cryogen ETHANE;60 second wait, 3-5 second blot, 597 filter paper, 0.5 second drain. Sample was supplemented with 10nm BSA-gold fiducials. 3.5ul of the mixture was double-side blotted. Resolution 9.60 Å

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

20 other PDB entries and 25 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name Q90VU7_9HIV1
Isoform
PDB entities 2
Chains and sequence ranges Author chain I; PDBConstruct 1–205; UniProt 2–206 Author chain K; PDBConstruct 1–205; UniProt 2–206 Author chain L; PDBConstruct 1–205; UniProt 2–206 Author chain N; PDBConstruct 1–205; UniProt 2–206

HLA class I histocompatibility antigen, A alpha chain

Homo sapiens

UniProt P04439

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 18 PDB declaration: octadecameric(18) Consistent with protein copy count Chain P; UniProt 334–365 Chain Y; UniProt 334–365 Mutation:T345S, S349G, G355S, C363A ADP-ribosylation factor 1 × 4 (P84077) Protein Nef × 4 (Q90VU7) AP-1 complex subunit beta-1 × 2 (Q10567) AP-1 complex subunit gamma-1 × 2 (P22892) AP-1 complex subunit mu-1 × 2 (P35585) AP-1 complex subunit sigma-3 × 2 (Q96PC3) GTP GUANOSINE-5'-TRIPHOSPHATE × 4 MG MAGNESIUM ION × 4 ELECTRON MICROSCOPY cryo-EM buffer:pH 7.2;HEPES/KOAc concentrated stocks are diluted to their final concentrations then pH'd to 7.2 with KOH prior to use in experiments. cryo-EM vitrification conditions:Cryogen ETHANE;60 second wait, 3-5 second blot, 597 filter paper, 0.5 second drain. Sample was supplemented with 10nm BSA-gold fiducials. 3.5ul of the mixture was double-side blotted. Resolution 9.60 Å

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

53 other PDB entries and 78 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name HLAA_HUMAN
Isoform
PDB entities 3
Chains and sequence ranges Author chain P; PDBConstruct 2–33; UniProt 334–365 Author chain Y; PDBConstruct 2–33; UniProt 334–365

AP-1 complex subunit beta-1

Homo sapiens

UniProt Q10567

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 18 PDB declaration: octadecameric(18) Consistent with protein copy count Chain A; UniProt 1–949 Chain B; UniProt 1–949 Mutation:K359R, E476K ADP-ribosylation factor 1 × 4 (P84077) Protein Nef × 4 (Q90VU7) HLA class I histocompatibility antigen, A alpha chain × 2 (P04439) AP-1 complex subunit gamma-1 × 2 (P22892) AP-1 complex subunit mu-1 × 2 (P35585) AP-1 complex subunit sigma-3 × 2 (Q96PC3) GTP GUANOSINE-5'-TRIPHOSPHATE × 4 MG MAGNESIUM ION × 4 ELECTRON MICROSCOPY cryo-EM buffer:pH 7.2;HEPES/KOAc concentrated stocks are diluted to their final concentrations then pH'd to 7.2 with KOH prior to use in experiments. cryo-EM vitrification conditions:Cryogen ETHANE;60 second wait, 3-5 second blot, 597 filter paper, 0.5 second drain. Sample was supplemented with 10nm BSA-gold fiducials. 3.5ul of the mixture was double-side blotted. Resolution 9.60 Å

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

19 other PDB entries and 19 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name AP1B1_HUMAN
Isoform
PDB entities 4
Chains and sequence ranges Author chain A; PDBConstruct 1–949; UniProt 1–949 Author chain B; PDBConstruct 1–949; UniProt 1–949

AP-1 complex subunit gamma-1

Mus musculus

UniProt P22892

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 18 PDB declaration: octadecameric(18) Consistent with protein copy count Chain E; UniProt 1–595 Chain G; UniProt 1–595 Not recorded ADP-ribosylation factor 1 × 4 (P84077) Protein Nef × 4 (Q90VU7) HLA class I histocompatibility antigen, A alpha chain × 2 (P04439) AP-1 complex subunit beta-1 × 2 (Q10567) AP-1 complex subunit mu-1 × 2 (P35585) AP-1 complex subunit sigma-3 × 2 (Q96PC3) GTP GUANOSINE-5'-TRIPHOSPHATE × 4 MG MAGNESIUM ION × 4 ELECTRON MICROSCOPY cryo-EM buffer:pH 7.2;HEPES/KOAc concentrated stocks are diluted to their final concentrations then pH'd to 7.2 with KOH prior to use in experiments. cryo-EM vitrification conditions:Cryogen ETHANE;60 second wait, 3-5 second blot, 597 filter paper, 0.5 second drain. Sample was supplemented with 10nm BSA-gold fiducials. 3.5ul of the mixture was double-side blotted. Resolution 9.60 Å

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

25 other PDB entries and 31 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name AP1G1_MOUSE
Isoform
PDB entities 5
Chains and sequence ranges Author chain E; PDBConstruct 1–595; UniProt 1–595 Author chain G; PDBConstruct 1–595; UniProt 1–595

AP-1 complex subunit mu-1

Mus musculus

UniProt P35585

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 18 PDB declaration: octadecameric(18) Consistent with protein copy count Chain J; UniProt 1–423 Chain M; UniProt 1–423 Not recorded ADP-ribosylation factor 1 × 4 (P84077) Protein Nef × 4 (Q90VU7) HLA class I histocompatibility antigen, A alpha chain × 2 (P04439) AP-1 complex subunit beta-1 × 2 (Q10567) AP-1 complex subunit gamma-1 × 2 (P22892) AP-1 complex subunit sigma-3 × 2 (Q96PC3) GTP GUANOSINE-5'-TRIPHOSPHATE × 4 MG MAGNESIUM ION × 4 ELECTRON MICROSCOPY cryo-EM buffer:pH 7.2;HEPES/KOAc concentrated stocks are diluted to their final concentrations then pH'd to 7.2 with KOH prior to use in experiments. cryo-EM vitrification conditions:Cryogen ETHANE;60 second wait, 3-5 second blot, 597 filter paper, 0.5 second drain. Sample was supplemented with 10nm BSA-gold fiducials. 3.5ul of the mixture was double-side blotted. Resolution 9.60 Å

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

22 other PDB entries and 29 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name AP1M1_MOUSE
Isoform
PDB entities 6
Chains and sequence ranges Author chain J; PDBConstruct 1–423; UniProt 1–423 Author chain M; PDBConstruct 1–423; UniProt 1–423

AP-1 complex subunit sigma-3

Homo sapiens

UniProt Q96PC3

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 18 PDB declaration: octadecameric(18) Consistent with protein copy count Chain O; UniProt 1–154 Chain S; UniProt 1–154 Not recorded ADP-ribosylation factor 1 × 4 (P84077) Protein Nef × 4 (Q90VU7) HLA class I histocompatibility antigen, A alpha chain × 2 (P04439) AP-1 complex subunit beta-1 × 2 (Q10567) AP-1 complex subunit gamma-1 × 2 (P22892) AP-1 complex subunit mu-1 × 2 (P35585) GTP GUANOSINE-5'-TRIPHOSPHATE × 4 MG MAGNESIUM ION × 4 ELECTRON MICROSCOPY cryo-EM buffer:pH 7.2;HEPES/KOAc concentrated stocks are diluted to their final concentrations then pH'd to 7.2 with KOH prior to use in experiments. cryo-EM vitrification conditions:Cryogen ETHANE;60 second wait, 3-5 second blot, 597 filter paper, 0.5 second drain. Sample was supplemented with 10nm BSA-gold fiducials. 3.5ul of the mixture was double-side blotted. Resolution 9.60 Å

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

19 other PDB entries and 19 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name AP1S3_HUMAN
Isoform
PDB entities 7
Chains and sequence ranges Author chain O; PDBConstruct 1–154; UniProt 1–154 Author chain S; PDBConstruct 1–154; UniProt 1–154

The page prioritizes protein identity, the current assembly, associated components, oligomeric state and cross-PDB links. Chain mapping and sequence ranges are retained as data evidence. Internal IDs, import timestamps and assembly operation expressions are maintenance fields and are not shown here.

SAXS scattering curve SAXS Profile

SAXS profile for 8d4d

P(r) Distance Distribution P(r) Distribution

P(r) distribution for 8d4d
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2. Structure Basics 2. Structure Basics

Entry ID entry_id8d4d
Deposition date deposition_date2022-06-01
Structure title titlegamma-Arf1 mediated dimeric assembly of AP-1, Arf1, Nef complex within lattice on MHC-I lipopeptide incorporated narrow membrane tubes
Keywords keywordsnef, AP, trafficking, PROTEIN TRANSPORT; PROTEIN TRANSPORT
Experimental Method methodELECTRON MICROSCOPY

3. SAXS Parameters (CRYSOL theoretical calculation) 3. SAXS Parameters (CRYSOL)

Radius of gyration Rg (Guinier) rg_guinier75.81
Radius of gyration Rg (electron density) rg_electron76.41
Forward intensity I(0) i02392530000.00
Molecular weight molecular_weight253000.0 kDa
Excluded volume excluded_volume248130 ų
Envelope volume envelope_volume842980 ų
Hydration-shell volume shell_volume97583 ų
Envelope diameter envelope_diameter249.1
Shell Rg shell_rg65.74
Envelope Rg envelope_rg73.16
Shape Rg shape_rg76.42
Total Rg total_rg76.22
Total atoms total_atoms18028
Residues n_residues4474
Spherical-harmonic order n_harmonics20
q range q_range— – 0.5000 −1
Data points n_points101
Shell type shell_typedirectional
Solvent electron density solvent_density0.3340 e/ų
Shell contrast contrast_shell0.0300 e/ų
CRYSOL version crysol_version4.1.3

4. P(r) Distance Distribution (GNOM inversion) 4. P(r) Analysis (GNOM)

Maximum dimension Dmax dmax219.8
Rg (real space) rg_real76.11
Rg uncertainty (real space) rg_real_error1.42
I(0) (real space) i0_real2.3910e+09
I(0) uncertainty (real space) i0_real_error4.6740e+07
Rg (reciprocal space) rg_reciprocal73.95
I(0) (reciprocal space) i0_reciprocal2381000000.0000
Solution quality estimate total_estimate0.8275
Solution quality rating solution_quality GOOD a GOOD solution
P(r) peaks n_peaks2
Primary peak position r_peak_primary62.8
Skewness Skewness skewness0.402
Kurtosis Kurtosis kurtosis-0.758
Angular range angular_range— – 0.1050 −1
Current regularization parameter α current_alpha0.0034
Highest regularization parameter α highest_alpha130700000.0000
Real-space data points n_real_points22
GNOM version gnom_version4.1.3
Quality Criteria quality_criteria AN1: 0.000; Oscil: 0.939; Stabil: 0.999; Sysdev: 1.000; Positv: 1.000; Valcen: 0.938; Smooth: 0.000

5. Crystallography and Experiment 5. Crystallography & Experiment

6. Entities and Polymers Entities & Polymers (9)

8. Citations (1)

9. Files and Curves (10)